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Background: Hepatitis B virus (HBV) infection is a serious public health problem in Africa and worldwide. Barbers are regularly in contact with the blood fluid of their clients, who could develop skin cuts and abrasions during shaving practices. There is scarcity of information about HBV among barbers in Cameroon. This study presents seroprevalence and associated risk factors for HBV infection among barbers and their clients in two cities (Yaounde and Douala) in Cameroon. Methods: Three millilitres of venous blood was collected into EDTA tubes from each of the 262 consenting participants. Plasma was obtained and transported to the Military Health Research Centre (CRESAR) for analysis. Sociodemographic data were collected using a structured questionnaire. The prevalence of Hepatitis B surface antigen (HBsAg) was determined by a rapid diagnostic test (DiaSpot HBsAg) and confirmed with an ELISA sandwich test. Data were analysed using SPSS, version 21. Results: Thirty-three participants tested positive, giving an overall prevalence of 12.6%. A seroprevalence of 15.0% and 10.6% was obtained for barbers and their clients respectively. In Yaounde, the frequency of HBsAg among barbers and their clients was 14.9% and 10.4% respectively, while in Douala it was 15.4% and 11.1% respectively. However, no statistically significant difference was observed in these groups. There was also no statistically significant association between the knowledge of the modes of HBV transmissions and HBV status. Conclusion: The present study reveals that the rate of HBsAg among the study population is high in Cameroon and most people are not still properly sensitised regarding prevention measures.